Study On The Analysis Method, Process Of Extraction And Purification Of Camptothecin

In this paper, a process of extraction and purification of Camptothecin from Camptotheca acuminata decne leaves had been studied systematically, the process courses, suitable for extraction and purification, had been explored.A sample reversed-phase high-performance liquid chromatography method (HPLC) was developed for determination of Camptothecin in Camptotheca acuminata decne leaves by extraction of Saxhlet. The samples were extracted by Saxhlet method at 80℃for 6 hours. The SinoChrom ODS-BP analytical column was used. The mobile phase of acetonitrile and water (40/60,v/v) was used to analysis Camptothecin at the wavelenth of 254nm. The method had a good linearity from the concentration of 3.55μg mL^-1 to 71μg mL^-1 with the recovery of 95.4% and the RSD of 2.52%.The process of extraction of soakage, refluence and NaOH aq. were studied for the extraction of Camptothecin from Camptotheca acuminata decne leaves. The separation and purification conditions were optimized by single factor and orthogonal experiment design. The optimal conditions of soakage extraction were as follows: concentration of methanol was 65%, extraction time was 48 hours, and solid to liquid rate was 1:8. And the average yield of Camptothecin was 0.31896o; The optimal conditions of refluence extraction were as follows: concentration of methanol was 65%, extraction time was one hour, solid to liquid rate was 1:10, and extraction times were once. The average yield of Camptothecin was 0.490%o. By extraction of NaOH aq. which was offered by WANG Yang etc, the average yield of Camptothecin was 0.490960. It was found that the refluence extraction method had less extraction time, simpler, higher extraction rate and further stability than other method.By performance of static absorption and adsorption experiment, resin of AB-8 was selected as the absorption carrier from five different styles of macroporous resins. The static absorption quantity of AB-8 resin could reached 16.68mg g^-1 dry resin and its static adsorption rate was 92.4% by eluting with 95% ethanol. We sublimated the rough extraction fluid by macroporous resin of AB-8 and confirmed the conditions of absorption and adsorption of the resin. The optimal absorption conditions were as follows: the initialize concentration was 0.175mg mL^-1, rough extraction fluid need not adjust its pH, absorption velocity of flow was 2BVh-1 and balance time of absorption was five hours. Camptothecin could be desorbed by using 95% ethanol at velocity of flow of 1 BV h^-1, and the elution rate could reach 83.1% at the elution quantity of 8BV. The purity of Camptothecin was reached 7.43%. The paper also analyzed the effect of usage times of resin on adsorption efficiency. It showed that absorption rate of the resin approximately reduced 30% when new resin was absorbed one time only. Therefore, we suggest that AB-8 macro porous resin should be regenerated systematically after it was used once.